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Here, we demonstrate that G\u03b1, an essential downstream component of GPCR signaling, undergoes a spatial shift from the plasma membrane to the endolysosomal compartment upon its constitutive activation. Using a genome-wide CRISPR screen, we identify the adaptor protein AP-3 as the essential mediator of this trafficking event. Loss of AP-3-dependent recruitment impairs G\u03b1-driven signaling and proliferation in uveal melanoma cells harboring oncogenic G\u03b1 mutations. We further identify a highly evolutionarily conserved AP-3 binding motif in all G\u03b1 proteins, present from yeast to humans. Disrupting this site causes G\u03b1 mislocalization and potently suppresses tumor growth and metastasis in vivo. Conversely, tethering G\u03b1 to the endolysosomal membrane is sufficient to rescue oncogenic signaling. Our findings reveal that endolysosomal recruitment is a fundamental, conserved requirement for G\u03b1 activity, uncovering a spatial vulnerability that may be exploited to target dysregulated G-protein signaling in human disease.<\/jats:p>","DOI":"10.1073\/pnas.2615774123","type":"journal-article","created":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T17:36:57Z","timestamp":1784741817000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":1,"title":["Oncogenic G\u03b1 signaling requires AP-3-dependent recruitment to the endolysosomal compartment"],"prefix":"10.1073","volume":"123","author":[{"given":"Megha","family":"Shettigar","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02jzgtq86","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medical Oncology, Dana-Farber Cancer 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